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人X染色体专性DNA片段的分离和鉴定

ISOLATION AND CHARACTERIZATION OF HUMAN X CHROMOSOME DNA SEGMENTS

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【作者】 张冬梅秦世真邱信芳蒋左庶薛京伦

【Author】 Zhang Dongmei;Qin Shizhen;Qiu Xinfang;Jiang Zuoshu;Xue Jinglun, (Institute of Genetics)

【机构】 复旦大学遗传学研究所复旦大学遗传学研究所

【摘要】 用人、鼠(CHO-K1)细胞总DNA为探针,对人X染色体DNA文库进行了三轮筛选。单拷贝DNA顺序的检出率为1.45%,即每筛选100个文库噬茵斑形成单位(plaque forming unit,pfu)可得到一二个单拷贝顺序。其中DXFD52,71,73,75分别与一组含人X染色体及不合人X染色体的人/鼠杂种细胞DNA进行Southern杂交后发现,DXFD52,73,75含人X染色体专性DNA顺序。结果表明,人X染色体DNA文库是获取X专性DNA片段的丰富来源。

【Abstract】 Random single-copy DNA segments were isolated from Los Alamos human Xchromosome library. The library was screened three times for nonrepetitive DNAinserts by plaque in situ hybridization to total human and mouse DNA. Out of 100plaque forming units (pfu), one to two single-copy recombinants, were obtained.DNAs from clones containing single-copy inserts were labeled by nick translationand Southern hybridization to normal human, and mouse DNA, and to a panel ofcell hybrid DNAs either retaining X chromosome or not retaining X chromosome.Human single-copy DNA segments isolated were all localized on X chromosome.Three segments, namely DXFD 52, DXFD 73, DXFD 75 were mapped to X chro-mosome. These segments will be further precisely localized to regions on X chro-mosome by in situ chromosomal hybridization.

【基金】 国家自然科学基金
  • 【文献出处】 复旦学报(自然科学版) ,Journal of Fudan University , 编辑部邮箱 ,1989年04期
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